The invention discloses a chitosan/arginine-glycine-aspartic acid (RGD) three-dimensional porous microcarrier, a preparation method thereof and application of the chitosan/RGD three-dimensional porous microcarrier to the culture of liver cells. The chitosan/RGD three-dimensional porous microcarrier is prepared from chitosan and RGD. The preparation method comprises the following steps of: allowing a mixed solution of the chitosan and the RGD to pass through a high-voltage pulse microcapsule forming instrument, dripping into a sodium polyphosphate solution in which the RGD is dissolved under the action of high pressure so as to form microspheres, preparing holes through freeze-drying, and finally forming the chitosan/RGD porous microcarrier with a three-dimensional structure. The aperture of the chitosan/RGD three-dimensional porous microcarrier is 20 to 50mm, the particle size is 400 to 900mm, and the porosity is 91 percent; meanwhile, by the adding of the RGD, the biocompatibility of the chitosan is effectively improved, and the liver cells can grow on the microcarrier in a high-density and high-activity way. The process is simple and easy to operate.